Yangtze University, Jingzhou, 434000, People's Republic of China; Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People's Republic of China.
J Virol Methods. 2020 May;279:113841. doi: 10.1016/j.jviromet.2020.113841. Epub 2020 Feb 24.
Japanese Encephalitis (JE) is an acute infectious disease that threatens both human and pig populations throughout Asia. JE is caused by the Japanese Encephalitis Virus (JEV), of which genotype III (GIII) had been the most prevalent strain throughout Asia, but recent studies have shown that genotype I (GI) has replaced GIII as the predominant version. Pigs and mosquitoes play a primary role in JEV transmission. However, a method for the rapid differentiation between JEV G I and G III remains unavailable. This study aimed to establish a rapid JEV genotyping method using novel duplex TaqMan RT-qPCR assay.specific primer and probes located in the PrM/M gene that were able to specifically differentiate GI and GIII JEV, was selected as the duplex TaqMan RT-qPCR target.The specificity, sensitivity and reproducibility test of this assay were validated. The sensitivity of the assay was 10 genomic RNA copies for both GI and GIII JEV in field mosquito and pig samples,and more sensitive than the current methods. In addition, the novel assay can be completed in less than 1 h. Therefore, This duplex TaqMan RT-qPCR assay is a promising tool for rapid differential detection and epidemiology of GI and GIII JEV strains in China. The results showed that co-circulation of GI and GIII infections with GI infection being more prevalent in pigs or mosquitoes in eastern China.
日本脑炎（JE）是一种急性传染病，威胁着整个亚洲的人类和猪群。JE 是由日本脑炎病毒（JEV）引起的，其中基因型 III（GIII）在整个亚洲一直是最流行的菌株，但最近的研究表明，基因型 I（GI）已取代 GIII 成为主要流行株。猪和蚊子在 JEV 传播中起主要作用。然而，快速区分 JEV GI 和 GIII 的方法仍然不可用。本研究旨在建立一种使用新型双重 TaqMan RT-qPCR 检测法快速鉴定 JEV 基因型的方法。选择位于 PrM/M 基因中的新型特异性引物和探针，能够特异性区分 GI 和 GIII JEV，作为双重 TaqMan RT-qPCR 检测的靶标。对该检测方法的特异性、敏感性和重复性进行了验证。该检测方法对田间蚊子和猪样本中的 GI 和 GIII JEV 的灵敏度均为 10 个基因组 RNA 拷贝，比目前的方法更灵敏。此外，该新方法可在 1 小时内完成。因此，这种双重 TaqMan RT-qPCR 检测法是一种快速区分和检测中国 GI 和 GIII JEV 株的有前途的工具。结果表明，GI 和 GIII 感染同时流行，在中国东部的猪或蚊子中，GI 感染更为普遍。
